International Journal of Advanced Research in Chemical Science (IJARCS)

Volume 5, Issue 2, 2018, PP 6-8

ISSN No. (Online) 2349-0403

DOI: http://dx.doi.org/10.20431/2349-0403.0502002

www.arcjournals.org

International Journal of Advanced Research in Chemical Science (IJARCS) Page | 6

Chemical Analysis of Hibiscus Rosa Sinenses-L

Bhawna Pandey1, S. K. Nigam

2, S. S. Paihar

1, Arvind Prasad Dwivedi

1Department of Chemistry, Govt. Girls P. G. College, Rewa-486001 (M.P.) India

2Govt. College Rampur Naikin, District Sidhi (M.P.) India

3Department of Chemistry, Govt. Sanjay Gandhi Smrati Auto., P.G., College Sidhi M.P

1. INTRODUCTION

Hibiscus a genus of flowering plants in malvaceae family contains 100 species that are native to warm

grown in subtropical and tropical regions throughout India. It includes both woody shrubs and small

trees. The flowers are large and of red and white colours. It is typically found almost everywhere. The

red Hibiscus is the flower of the Hindu goddess Kali in Bengal India and offered goddess and Lord Ganesha in Hindu worship.

It has a long history as for health benefits are concerned, its tea is popular lowers blood pressure, decrease the concentration of LDL cholesterol. It is used as hair oil, paste, shampoo plus conditions,

cosmetic skin care, anti-solar agent, notably in reduction of triglycerides and shows as anti-oxidant

effect.2 Further studies are needed to demonstrate clinical significance. The suitable analytical

procedure for the analysis3 of phytoconstituents such as chlorophylls, amino acids, lignans is of

technical importance. In present investigation, we report here chromatographic fluorimetric,

electrophoretic and other physical methods that provide good results of closely related constituents in different varieties of HRS medicinal plants. The various aspects of the analysis have been described.

2. EXPERIMENTAL

All the chemicals used in the study were of analytical grade. The solvents employed were distilled and

their identities confirmed with the authentic standard methods. The biomass as an, electrolyte was isolated after performing various primary processes such as plucking, crushing and extraction of

leaves from HRS medicinal plants collected as pooled samples.

The maximum content of phytoconstituents of HRS1, HRS2, and HRS3 was obtained by extracting

with methanol separately. The extracts were filtered, dried under vacuum and made up to 1 ml with

CH3OH prior to chromatographic analysis.4,5

The ascending paper chromatography was performed for measuring Rf values of the samples under a flow of nitrogen gas at experimental conditions of

temperature and relative humidity. Several others analytical techniques6,7

involving fluorimetry,8 UV

spectrometry9, electrophoresis etc. have been described for the determination of lignans and other

compounds10

of the system. The pH, conductivity of biomass of medicinal plants have been digitally

measured by pH and conductivity meter while the number of cells in sap is counted by research

microscope.11

3. RESULTS AND DISCUSSION

The natural chlorophyll-b and chlorophyll-a along with xanthophyll and carotenes in two yellow and

two green pigments were calculated from the extracts of the HRS1, HRS2, and HRS3. The results of Rf

Abstract: The chemical analysis of bio-sap of medicinal plants Hibiscus Rosa Sinenses-L (HRS) of

malvaceae family have been performed chromatographically and documented. The work is restricted to the

identification of pigments, and free amino acids. Various physical parameters related to analysis were

determined. Electrophoretic method was employed to measure the relative motion of ions of amino acids. The

fluorescent and UV spectroscopic methods were used for the active ingredient’s analysis.

Keywords: Identification, ingredients, diffusion, relative mobility, extraction.

*Corresponding Author: Bhawna Pandey, Department of Chemistry, Govt. Girls P. G. College, Rewa-

486001 (M.P.) India

Chemical Analysis of Hibiscus Rosa Sinenses-L

International Journal of Advanced Research in Chemical Science (IJARCS) Page | 7

values of three determinations obtained by ascending paper chromatography analysis, were comparable with those of Rf reference values (Table 1). The chloroplast has been observed

accompanying electron flow and formation of ATP during photic excitation (Phytosynthesis). The

energy of photic excitation is converted into ATP and reduction of NADP to NADPH and H+.

Swelling and shrinking of chloroplast have been observed during photosynthesis. That is why; HRS3

possesses much higher value in comparison to HRS2 and HRS1 respectively. The nine amino acids

were detected chromatographically from the extracts of the samples of HRS and their Rf values are

recorded in (Table 2). The Rf values facilitated the identification by ninhydrin locating reagent.

The Rf values of phyllanthin and hypophyllanthin from other closely related lignans were also determined as 0.17 and 0.20 respectively. The chromatographic separation was also carried out for

carbohydrates (Table 3). The fluorescent analysis of the samples under investigation using UV light

gave different colors were also analyzed. Various physical parameters such as pH, conductivity,

density and surface tension have also been determined (Table 4).

The relative mobility of amino acids (known and unknown) was studied in samples of variants HRS by electrophoretic method at pH 1.81 and 5.2 respectively (Table 5 and 6). It was observed that

electrophoresis at low voltage is not usually used/ permissible to separate low molecular weight

amino acids because of diffusion but it is easier to illustrate the relationship between charge and pH

with amino acids than with proteins or other macromolecules. With exception of other amino acids was not detected in any variants of HRS analyzed.

Table1. Identification of pigments for Hibiscus Rosa Sinenses-L., by chromatographic method

S. No. Pigments Colour Rf values

HRS1 HRS2 HRS3

1. Chlorophyll-b Green 0.26 0.33 0.37

2. Chlorophyll-a Blue green 0.35 0.29 0.30

3. Xanthophyll Yellow brown 0.59 0.64 0.75

4. Carotene Yellow 0.85 0.87 0.91

Table2. Chromatographic separation of Amino acids

S.

No.

Pigments Colour Rf values References

values HRS1 HRS2 HRS3

1. Alanine

(α-amino propionic acid)

Light brown

purple

0.35 0.39 0.41 0.42

2. Aspartic acid

(α-amino succinic acid)

Blue 0.11 0.15 0.13 0.19

3. Glycine (amino acetic

acid)

Deep bluish

purple

0.49 0.52 0.47 0.58

4. Leucine

(α-amino isocaproic acid)

Light purple

bluish

0.84 0.79 0.84 0.88

5. Proline (α-carboxylic acid)

Yellowish violet

0.77 0.89 0.78 0.94

6. Phenyl analine Blue brown 0.11 - 0.13 0.85

7. Tyrosine

(α-amino-β-cp-hydroxy

phenyl) propionic acid

Purple

yellowish

violet

0.76 0.77 0.75 0.82

8. Threonine

(δ –amino-β-hydroxyl-n-

butyric acid

Blue light 0.71 0.78 0.79 0.82

9. Valine Blue 0.66 0.69 0.68 0.73

Table3. Chemical analysis of carbohydrates by ascending paper chromatographic method

S. No. Sugar Rf values References

Rf values HRS1 HRS2 HRS3

1. D-fructose 0.18 0.20 0.17 0.21

2. D-Lactose 0.71 0.72 0.69 0.70

3. D-glucose 0.098 0.091 0.083 0.10

4. L-Sorbose 0.22 0.19 0.23 0.20

5. R-affinose 0.088 0.093 0.07 0.09

6. D-galactose 0.15 0.18 0.16 0.17

Chemical Analysis of Hibiscus Rosa Sinenses-L

International Journal of Advanced Research in Chemical Science (IJARCS) Page | 8

Table4. Physical parameters for medicinal plant Hibiscus Sinenses-L

S. No. System pH Density

Gram/cm3

Surface tension

dynes/cm

Conductivity

Siemen(s)

1. HRS1 5.51 0.891 41.1 2.73 × 10-3

2. HRS2 5.48 0.896 41.8 2.69 × 10-3

3. HRS3 5.47 0.906 40.8 2.71 × 10-3

Table5. Relative Mobilities of known Amino acids (Electrophoretic method)

S. No. Compound Relative mobility in electrode (mm)

Ia 2b

1. Alanine -100 -100

2. Glycine -116 -

3. Leucine -77 -78

4. Threonine -75 -73

a. 2.5% (W/V) HCOOH-7.8 (W/V) CH3COOH, Mobility relative to alanine pH 1.87,

100 V/Cms;

b. 2% (W/V) HCOOH- 20% (W/V) CH3COOH- 0.4 mm Cd acetate pH 1.84,

100 V/Cm, Mobility relative to alanine.

Table6. Relative Mobilities of unknown (observed values) of Amino acid at pH 5.2 (Electrophoretic method)

S. No. Compound Relative mobility in electrode (mm)

Ia 2b

1. Alanine -102 -102

2. Glycine -116 -

3. Leucine -79 -78

4. Threonine -78 -78

4. CONCLUSION

The analysis of lignans and pigments by modern techniques would be very helpful for others

medicinal plants too. The study may be extended for use an anti-oxidant, and in practicing traditional system of generic medicines, as-well-as cosmetics and detergents industries.

REFERENCES

[1] Harvey, A.L. Drug Discovery Today, 2009, 13: (19-20), 849-901.

[2] Khatoon, S., Rai, V., Rawat, A.K.S., and Mehrotra, S., J. Ethnophamac, 2006, 104: 79-86.

[3] Anupama, P., Ayurveda Herb. 2013, 1: 6.

[4] Tripathi, Arvind, K., Verma, Ram K., Gupta, Anil K., Madan, M., and Suman, Khanuja P.S., Phytochem.

Anal., 2006, 17, 394-397.

[5] Frey, H.P. and Zieloff, K. Qualitative and Quantitative Chromatography, Grund lagen and Praxis, VCO,

1993.

[6] Wang, C.Y., and Lee, S.S., Phytochem. Anal., 2005, 16: 120-126

[7] Shrivastava, S., Gupta, M.M., Verma, R.K., and Kumar, S., J. Assoc. off Anal Chem. (Int.), 2000, 83:

1484-1488.

[8] Lakowiczj, J.R., Principles of Fluorescence spectroscopy, Kluwer Academic Plenum Publishers, 1999.

[9] Vogel, I. Elementary, practical organic chemistry, Pearson, Education Publication, 2010.

[10] Bunkar, Alka Rani, Int. J. Chem. Studies, 2017, 5(1): 09-11.

Citation: B. Pandey, "Chemical Analysis of Hibiscus Rosa Sinenses-L", International Journal of Advanced

Research in Chemical Science (IJARCS), vol. 5, no. 2, pp. 6-8, 2018. http://dx.doi.org/10.20431/2349-

0403.0502002

Copyright: © 2018 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium,

provided the original author and source are credited.